A simple method for functionalization of cellulose membrane for covalent immobilization of biomolecules

Bora, Utpal; Krishnamoorthy, K.; Nahar, Pradip

doi:10.1016/j.memsci.2004.10.028

Cited by 50 publications

(30 citation statements)

References 21 publications

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“…The immobilized enzyme maintained 70% of its maximal activity in the pH range of 4-7, while free enzyme maintained 70% activity levels in the pH range of 5-6. The pH profile change in immobilized enzymes is very common and was confirmed by other authors [12,35,36]. It was found that the immobilized α-galactosidase was much more stable than free enzymes, indicating that free enzymes are more sensitive to the pH changes, proving that the enzyme stability in different pH concentrations was significantly improved by immobilization.…”

Section: Resultssupporting

confidence: 62%

Covalent Immobilization of α-Galactosidase from Penicillium griseoroseum and its Application in Oligosaccharides Hydrolysis

Falkoski

Guimarães

Queiroz

et al. 2008

Appl Biochem Biotechnol

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Partially purified alpha-Galactosidase from Penicillium griseoroseum was immobilized onto modified silica using glutaraldehyde linkages. The effective activity of immobilized enzyme was 33%. Free and immobilized alpha-galactosidase showed optimal activity at 45 degrees C and pH values of 5 and 4, respectively. Immobilized alpha-galactosidase was more stable at higher temperatures and pH values. Immobilized alpha-galactosidase from P. griseoroseum maintained 100% activity after 24 h of incubation at 40 degrees C, while free enzyme showed only 32% activity under the same incubation conditions. Defatted soybean flour was treated with free and immobilized alpha-galactosidase in batch reactors. After 8 h of incubation, stachyose was completely hydrolyzed in both treatments. After 8 h of incubation, 39% and 70% of raffinose was hydrolyzed with free and immobilized alpha-galactosidase respectively. Immobilized alpha-galactosidase was reutilized eight times without any decrease in its activity.

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Section: Resultssupporting

confidence: 62%

Covalent Immobilization of α-Galactosidase from Penicillium griseoroseum and its Application in Oligosaccharides Hydrolysis

Falkoski

Guimarães

Queiroz

et al. 2008

Appl Biochem Biotechnol

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“…1A). Light-induced attachment of the photolinker to the solid surface was confirmed by X-ray photoelectron spectroscopy in our earlier study, 23,28 which showed the presence of nitrogen and fluorine in the ratio of 2 : 1 in the activated surface. The labile fluoro group of the activated polystyrene is easily displaced by nucleophilic (such as thiol or amine) attack of the modified oligonucleotide and forms covalent linkage between activated surface and the modified oligonucleotide.…”

Section: Resultssupporting

confidence: 53%

Single-step covalent immobilization of oligonucleotides onto solid surface

2010

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In this communication, we report a simple, single-step method for covalent immobilization of oligonucleotide probes onto an activated polystyrene surface. Polystyrene surface is activated by a photolinker, 1-fluoro-2-nitro-4-azidobenzene (FNAB) in a photochemical reaction by UV light. Amine-modified oligonucleotide probe is covalently immobilized to the activated surface by displacing the labile fluoro group of the activated polystyrene surface by the primary amino group of the aminated oligonucleotide probe. Biotinylated complementary oligonucleotide target is hybridized with the immobilized probe. The hybridized target is detected by binding streptavidin-peroxidase followed by assaying the enzyme for colorimetric signal measurement. The covalently immobilized oligonucleotide probe shows high selectivity in subsequent hybridization processes with the complementary target and clearly discriminates with single-base mismatched oligonucleotide targets. The method can immobilize amine-or thiol-modified oligonucleotide onto FNAB-activated polymer surface. The method could be potentially useful for designing DNA based biosensors or biochips and also in microtiter plate based assay systems in a cost effective manner.

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“…Therefore, the immobilization led to a considerable increase of thermostability. Also, immobilized HRP on cellulose membrane retained full activity at least for 2 h at 60°C, whereas the activity of free enzyme was reduced to almost zero at the same temperature [36]. In the same manner, immobilized turnip peroxidase incubated at 60°C for 2 h retained 54% of the initial enzyme activity, whereas the soluble enzyme lost nearly 83% of the original activity under similar incubation conditions [9].…”

Section: Kmmentioning

confidence: 76%

Immobilization of Horseradish Peroxidase on Nonwoven Polyester Fabric Coated with Chitosan

Mohamed

Aly

Mohamed

et al. 2007

Appl Biochem Biotechnol

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The immobilization of horseradish peroxidase (HRP) on composite membrane has been investigated. This membrane was prepared by coating nonwoven polyester fabric with chitosan glutamate in the presence of glutraldehyde as a crosslinking agent. The physicochemical properties of soluble and immobilized HRP were evaluated. The soluble HRP lost 90% of its activity after 4 weeks of storage at 4 degrees C, whereas the immobilized enzyme retained 85% of its original activity at the same time. A reusability study of immobilized HRP showed that the enzyme retained 54% of its activity after 10 cycles of reuse. Soluble and immobilized HRP showed the same pH optima at pH 5.5. The immobilized enzyme had significant stability at different pH values, where it had maximum stability at pH 3.0 and 6.0. The kinetic properties indicated that the immobilized enzyme had more affinity toward substrates than soluble enzyme. The soluble and immobilized enzymes had temperature optima at 30 and 40 degrees C and were stable up to 40 and 50 degrees C, respectively. The stability of HRP against metal ion inactivation was improved after immobilization. Immobilized HRP exhibited high resistance to proteolysis by trypsin. The immobilized HRP was more resistant to inactivation induced by urea, Triton X-100, and organic solvents compared to its soluble counterpart. The immobilized HRP showed very high yield of immobilization and markedly high stabilization against several forms of denaturants that offer potential for several applications.

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A simple method for functionalization of cellulose membrane for covalent immobilization of biomolecules

Cited by 50 publications

References 21 publications

Covalent Immobilization of α-Galactosidase from Penicillium griseoroseum and its Application in Oligosaccharides Hydrolysis

Covalent Immobilization of α-Galactosidase from Penicillium griseoroseum and its Application in Oligosaccharides Hydrolysis

Single-step covalent immobilization of oligonucleotides onto solid surface

Immobilization of Horseradish Peroxidase on Nonwoven Polyester Fabric Coated with Chitosan

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